Method and compositions for treatment of cancers

ABSTRACT

A method to treat cancer uses ultrapheresis, refined to remove compounds of less than 120,000 daltons molecular weight, followed by administration of replacement fluid, to stimulate the patient&#39;s immune system to attack solid tumors. In the preferred embodiment, the patient is ultrapheresed using a capillary tube ultrafilter having a pore size of 0.02 to 0.05 microns, with a molecular weight cutoff of 120,000 daltons, sufficient to filter one blood volume. The preferred replacement fluid is ultrapheresed normal plasma. The patient is preferably treated daily for three weeks, diagnostic tests conducted to verify that there has been shrinkage of the tumors, then the treatment regime is repeated. The treatment is preferably combined with an alternative therapy, for example, treatment with an anti-angiogenic compound, one or more cytokines such as TNF, gamma interferon, or IL-2, or a procoagulant compound. The treatment increases endogenous, local levels of cytokines, such as TNF. This provides a basis for an improved effect when combined with any treatment that enhances cytokine activity against the tumors, for example, treatments using alkylating agents, doxyrubicin, carboplatinum, cisplatinum, and taxol. Alternatively, the ultrapheresis treatment can be combined with local chemotherapy, systemic chemotherapy, and/or radiation.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.09/699,003, filed Oct. 26, 2000, which is a continuation of U.S.application Ser. No. 09/316,226, now U.S. Pat. No. 6,231,536, filed May21, 1999, which is a continuation-in-part-of U.S. application Ser. No.09/083,307, now U.S. Pat. No. 6,620,382, filed May 22, 1998, each ofwhich is hereby incorporated by reference.

BACKGROUND OF THE INVENTION

The present invention is generally in the field of enhancing an immuneresponse, and particularly relates to the removal of inhibitors ofimmune mediators, in combination with anti-angiogenic compounds,cytokines, compounds inducing a procoagulant state, chemotherapeuticsand/or radiation.

Conventional cancer therapy is based on the use of drugs and/orradiation which kills replicating cells, hopefully faster than theagents kill the patient's normal cells. Surgery is used to reduce tumorbulk, but has little impact once the cancer has metastasized. Radiationis effective only in a localized area.

The treatments can in themselves kill the patient, in the absence ofmaintenance therapy. For example, for some types of cancer, bone marrowtransplants have been used to maintain the patient following treatmentwith otherwise fatal amounts of chemotherapy. Efficacy has not beenproven for treatment of solid tumors, however. “Cocktails” of differentchemotherapeutic agents and combinations of very high doses ofchemotherapy with restorative agents, for example, granulocytemacrophage colony stimulating factor (“GM-CSF”), erythropoietin,thrombopoetin, granulocyte stimulating factor, (“G-CSF”), macrophagecolony stimulating factor (“M-CSF) and stem cell factor (“SCF”) torestore platelet and white cell levels, have been used to treataggressive cancers. Even with the supportive or restrictive therapy,side effects are severe.

Other treatments have been tried in an attempt to improve mortality andmorbidity. Vaccines to stimulate the patient's immune system have beenattempted, but not with great success. Various cytokines, alone or incombination, such as tumor necrosis factor, interferon gamma, andinterleukin-2 (“IL-2”) have been used to kill cancers, but have notproduced cures. More recently, anti-angiogenic compounds such asthalidomide have been tried in compassionate use cases and shown tocause tumor remission. In animal studies, compounds inducing aprocoagulant state, such as an inhibitor of protein C, have been used tocause tumor remission. New studies have shown that inhibitors ofcytokine receptors, such as tumor necrosis factor receptors (“TNF-Rs”)which are released in a soluble form from tumor cells, in highconcentrations relative to normal cells, may restore the immune system'sattack on the tumor cells (Jablonska and Peitruska, Arch. Immunol. Ther.Exp. (Warsz) 1997, 45(5-6), 449-453; Chen, et al., J. Neuropathol. Exp.Neurol. 1997, 56(5), 541-550).

U.S. Pat. No. 4,708,713 to Lentz describes an alternative method fortreating cancer, involving ultrapheresis to remove compounds based onmolecular weight, which promotes an immune attack on the tumors by thepatient's own white cells.

Despite all of these efforts, many patients die from cancer; others areterribly mutilated. It is unlikely that any one therapy will beeffective to cure all types of cancer.

It is therefore an object of the present invention to provide a methodand compositions for treatment of solid tumors.

It is a further object of the present invention to provide a method andcompositions that does not involve non-selective, extremely toxic,systemic chemotherapy.

SUMMARY OF THE INVENTION

A method to treat cancer uses ultrapheresis, refined to remove compoundsof less than 120,000 daltons molecular weight, followed byadministration of replacement fluid, to stimulate the patient's immunesystem to attack solid tumors. In the preferred embodiment, the patientis ultrapheresed using a capillary tube ultrafilter or parallel platefilter having a molecular weight cutoff of 120,000 daltons, sufficientto filter at least one blood volume. The preferred replacement fluid isultrapheresed normal plasma. Alternatively, the patient is pheresed toselectively remove soluble receptor/inhibitors to soluble tissuenecrosis factor receptor-1 (“sTNFR-1”), soluble tissue necrosis factorreceptor-2 (“sTNFR-2”), soluble interleukin-2 receptor (“sIL-2R”),soluble interleukin-1 receptor (“sIL-1R”), soluble interleukin-6receptor (“sIL-6R”), or soluble interferon-gamma receptor(“sIFN-gammaR”). These can be removed by binding to the cytokine, anepitope thereof, or an antibody to the receptor. These can beimmobilized in the filter, in a column, or using other standardtechniques for binding reactions to remove proteins from the blood orplasma of a patient. The patient is preferably treated daily for threeweeks, diagnostic tests conducted to verify that there has beenshrinkage of the tumors, then the treatment regime is repeated.

The treatment is preferably combined with an alternative therapy, forexample, treatment with an anti-angiogenic compound, one or morecytokines such as TNF, gamma interferon, other interferons, or IL-2, ora procoagulant compound. The treatment increases the inflammationagainst tumors by allowing cytokines, such as TNF, to work effectively.This provides a basis for an improved effect when combined with anytreatment that enhances cytokine activity against the tumors, forexample, treatments using alkylating agents, doxyrubicin, carboplatinum,cisplatinum, and taxol, and other drugs which may be synergistic ineffect with “unblocked” cytokines. Alternatively, the ultrapheresistreatment can be combined with local chemotherapy, systemicchemotherapy, and/or radiation.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1 and 2 are schematics of the system for ultrapheresis.

DETAILED DESCRIPTION OF THE INVENTION

The methods and devices disclosed herein are useful for treatment ofpatients with cancer, immune-mediated disorders, chronic parasitism,some viral diseases, and other disorders characterized by elevatedlevels of TNF receptors or inhibitors to IL-2, IL-6, gamma interferon,or other pro-inflammatory signals as well as white cell activation.Examples demonstrate efficacy in treating cancer patients.

I. Ultrapheresis

A. Ultrapheresis System

1. Filters

The filter must be biocompatible, and suitable for contact with blood,without causing excessive activation of platelets or clotting. Deviceswill typically be either parallel plate filters or capillary membranefilters. These can be adapted from devices currently in use for kidneydialysis. The capillary membrane filters will typically have a surfacearea of between about 0.25 and 1 m² for use with children and betweenabout 1 and 3 m² for use with adults. The parallel plate filters willtypically have a surface area in the range from 0.1 and 2 cm²/ml ofblood to be filtered.

The filter membranes will typically be a biocompatible or inertthermoplastic such as polycarbonate, polytetrafluorethylene (Teflon®),polypropylene, ethylene polyvinyl alcohol or polysulfone, having a poresize typically of between 0.02 and 0.05 microns in a capillary membranefilter and of between 0.04 and 0.08 microns in a parallel plate filter.The actual pore size that yields the desired cutoff of approximately120,000 daltons is determined based on the fluid flow geometry, shearforces, flow rates, and surface area. The effective cutoff for acapillary membrane filter with a pore size of 0.03 microns is 120,000daltons, with a sieving coefficient of between 10 and 30%. This resultsin only a trivial amount of IgG being removed from the patient's blood.The filter membrane should be less than about 25 microns, preferablyless than about 10 microns, thick. The permeable membrane should notcause blood clotting or otherwise react with the blood.

A preferred membrane is one in which the pores are made by electronbeams directed perpendicularly to the surface because the size anddensity of the pores can be accurately controlled in this manner. Thepores are essentially circular in cross section so the effective poresize is the actual pore size. The effective pore size of ultrafilteredmedia having pores with non-circular cross sections shall be thediameter of a circular pore which will pass molecules or othercomponents of an equivalent size to the molecules or other componentswhich pass through the filter medium in question.

Suitable devices can be obtained from Asahi Chemical Company, Japan, andKuraray Co., Ltd, 1-12-39, Umeda, ite-ku, Osaka 530, Japan.

Staged filters can also be used, which have different pore sizes and/orgeometries or surfaces areas, to provide for a “staggered” removal ofmaterials from the blood. Alternatively, although not at this timepreferred, one can use differential centrifugation, to provide for anappropriate separation of blood components. Specific absorbing columnscan also be employed to selectively remove specific cytokine andcellular inhibitors from the filtered plasma so that the ultrafiltrate(treated plasma) can be returned to the patient.

2. Process Controls and Fluid Handling

The patient will typically be connected to the blood processing deviceusing standard intravenous tubing, with connections similar to thoseused for plateletpheresis, so that blood can be removed from the patientat one site and returned at another. The tubing is connected to a pumpthat controls the flow rate so that in the preferred embodiment oneblood volume (based on approximately 7% of the total body weight) isprocessed over a period of approximately 2½ hours. The filtrate is thenreturned from the filtration device to the patient at the second site.Standard microprocessor controls can be used to regulate the blood flow,for example, by monitoring the volume of the blood products beingremoved, in combination with flow rate monitors and pump speed.

The entire system should first be flushed with saline and then treatedwith an anticoagulant or anticlotting agent, such as sodium heparin, tobe sure that there are no locations within the system where bloodclotting can occur. Moreover, small amounts of anticoagulants should becontinually introduced into the blood stream directed to the ultrafilterto ensure than no clotting occurs during the filtration process. All ofthe surfaces of the system which come in contact with the blood andfluids which are infused into the patient must be sterilized prior tocommencing treatment.

FIG. 1 illustrates a system for ultrapheresis. Blood is removed from apatient by means of a venous catheter 10 with the distal lead 11 thereofdisposed in the superior vena cava 12 leading to the patient's heart 13.The blood passes through conduit 14 to a drip chamber 15 and then intopump 16 which controls the pressure of the blood to the separation unit17, preferably an ultrafilter as shown, through conduit 18. A pressuregauge 19 is provided on conduit 14 to continually monitor arterialpressure. A syringe pump 20 feeds an anti-clotting drug such as sodiumheparin to conduit 18 to prevent the clotting of blood in theultrafilter 17. In the ultrafilter 17 the blood stream passes over theultrafilter medium or membrane 21 under pressure. The blood fractionincluding the low molecular weight components passes through themembrane 21 and is discharged as permeate through conduit 22. Theretentate or treated blood containing the high molecular weightcomponents, which include whole blood cells and platelets, is dischargedinto conduit 23 which ultimately leads back to the patient. Volumetricpump 27 passes a controlled amount of permeate to a container 28 forcontainment and for measuring. Volumetric pump 30, which is preferablythe same type and capacity as pump 27, pumps replacement fluid from acontainer 31 to conduit 32, which directs the fluid to conduit 23 whereit mixes with the retentate or treated blood. The treated blood andother components are returned to the patient through venous catheter 34,the distal or discharge end of which is disposed in the brachiocephalicvein. The volumetric pumps 27 and 30 are preferably set either to pumpthe same total amount of fluid or to pump at the same rate, so that thesame volume of fluid which is removed from the patient's blood stream aspermeate is returned as replacement fluid. The blood stream in conduit23 is passed through filter 36 to remove clots or other debris from theblood stream. A drip chamber 37 ensures that no significant quantitiesof air enter the patient's blood stream. A pressure gauge 38 is providedto continually monitor venous blood pressure.

FIG. 2 illustrates another embodiment wherein blood removed from apatient is first passed through conduit 30 to a first ultrafilter 31 toselectively separate a blood fraction with components having molecularweights less than about 1,000,000 Daltons. The retentate from thisultrafiltration which contains the high molecular weight components isreturned through conduit 32 to the patient. The permeate from the firstultrafilter, 31 is passed through conduit 33 to a second ultrafiter 34where a blood fraction having a molecular weight below 30,000 is removedfrom the permeate stream from the first ultrafilter 31. The permeatefrom the second ultrafiter 34, which contains the very low molecularweight components such as salts and nutrients may be returned to thepatient through conduit 35. The retentate from the second ultrafilterwhich contains blocking factors, IgG immunoglobulins and othercomponents is discharged through conduit 36.

Blood should be pumped through the ultrafilter unit at sufficientpressure to cause the blood components having the immunosuppressiveeffects to pass through the filter but at a velocity which will notexcessively shear or otherwise damage the blood cells passing over themembrane. Generally it has been found that the ratio of the area of themembrane to the amount of blood treated per hour should be within about0.1 to 2 cm/mL. Differential pressure across the membrane should rangefrom about 2 to 20 mM Hg.

3. Replacement Fluids

The patient must receive replacement fluids following filtration. Thepreferred replacement fluid is ultrapheresis normal plasma, for example,expired plasma obtained from the Red Cross, which has been filteredusing the same filter as used to treat the patient. Alternatively, thepatient can be administered normal albumin, or fresh frozen plasmadiluted with saline.

II. Treatment with Adjuvant Therapies

Standard ultrapheresis is conducted over a period of time until apositive indication is observed. This is typically based on diagnostictests which show that there has been some reduction in tumor size orwhich suggests tumor inflammation. The patient is preferably treateddaily for three weeks, diagnostic tests conducted to verify that therehas been shrinkage of the tumors and/or inflammation, then the treatmentregime is repeated.

Surgical (or vacuum) removal of necrotic material may be required priorto or during treatment to avoid toxicity associated with high tumorburden.

This procedure has been demonstrated to cause a significant response(greater than 50% reduction in size of tumors) in a variety of solidtumors in approximately 50% of patients who have failed all othertreatment modalities. A tumor specific inflammatory response provoked byultrapheresis has been documented in approximately 75% of patients inmetastatic melanoma clinical trials. This inflammation is characterizedby redness, swelling, warmth, and tenderness and is confined to tumorsonly. There has been no associated injury to non-cancerous tissue. Thistumor specific inflammatory response has led to a 50% or greaterreduction in the size of tumors in 50% of patients studied so far.Clinical trials have also demonstrated a 44% major reduction of tumormetastases in human breast cancer and prostate cancer.

Tumor specific inflammation has been observed in patients withmetastatic colon cancer, ovarian cancer, lung cancer, head and neckcancer, cervical and endometrial cancers. In some cases, thisinflammation has been followed by significant tumor regressions in eachtumor type. The significance of response in such diverse tumor typesstrongly suggests that ultrapheresis modifies the patient's response tothe tumor in favor of successful immunologic control of the tumor. Typesof tumors that are particularly sensitive to the ultrapheresis includeepithelial tumors, sarcomas, melanomas and glioblastomas.

It would clearly be advantageous to cause complete remissions. Based onthe presumed mechanism that the process is removing immune inhibitorsproduced by the tumors, especially inhibitors of cytokines and otherimmune mediators, it is possible to treat the patients with adjuvant orcombination therapies, that enhance the results achieved with theultrapheresis. TNF-alpha and beta receptors are thought to beparticularly important immune inhibitors. Therefore, compounds whichenhance TNF activity are particularly preferred. These includeanti-angiogenic compounds, such as thalidomide, procoagulant compounds,cytokines and other immunostimulants. Standard chemotherapeutic agentsand/or radiation can also be used with the ultrapheresis.

A. Anti-Angiogenic Compounds

Any anti-angiogenic compound can be used. Exemplary anti-angiogeniccompounds include O-substituted fumagillol and derivatives thereof, suchas TNP-470, described in U.S. Pat. Nos. 5,135,919, 5,698,586, and5,290,807 to Kishimoto, et al.; angiostatin and endostatin, described inU.S. Pat. No. 5,290,807, 5,639,725 and 5,733,876 to O'Reilly;thalidomide, as described in U.S. Pat. Nos. 5,629,327 and 5,712,291 toD'Amato; and other compounds, such as the anti-invasive factor, retinoicacid, and paclitaxel, described in U.S. Pat. No. 5,716,981 to Hunter, etal., and the metalloproteinase inhibitors described in U.S. Pat. No.5,713,491 to Murphy, et al. Thalidomide is administered once daily, 200mg orally.

B. Procoagulant Compounds

Protein C is a vitamin K-dependent plasma protein zymogen to a serineprotease. Upon activation it becomes a potent anticoagulant. Activatedprotein C acts through the specific proteolysis of the procoagulantcofactors, factor VIIIa and factor Va. This activity requires thepresence of another vitamin K-dependent protein, protein S, calcium anda phospholipid (presumably cellular) surface. As described in Hemostasisand Thrombosis: Basic Principles and Clinical Practice 2nd Ed., Colman,R. W., et al., p. 263 (J.B.Lippincott, Philadelphia, Pa. 1987), proteinC circulates in a two-chain form, with the larger, heavy chain bound tothe smaller light chain through a single disulfide link. Protein C isactivated to activated protein C (APC). Thrombin is capable ofactivating protein C by the specific cleavage of the Arg¹²-Leu¹³ bond inthe heavy chain. In vivo, in the presence of physiologicalconcentrations of calcium, the rate of this activation is enhanceddramatically when thrombin is bound to the endothelial cell cofactor,thrombomodulin. Matschiner, et al., Current Advances in Vitamin KResearch, pp. 135-140, John W. Suttie, ed. (Elsevier Science PublishingCo., Inc. 1988) have further reviewed the role of the Vitamin Kdependent proteins in coagulation.

Blockage of the natural anticoagulant pathways, in particular theprotein C pathway, uses the natural procoagulant properties of the tumorto target the tumor capillaries for microvascular thrombosis, leading tohemorrhagic necrosis of the tumor, as described in U.S. Pat. No.5,147,638 to Esmon, et al. Examples of such compounds includeanti-protein C and anti-protein S.

C. Cytokines

The biologic activity and clinical effectiveness of pro-inflammatorycytokines is augmented by ultrapheresis in the patient with cancer andother states of acquired immune tolerance Specifically, both TNF alphaand TNF beta, in doses of between approximately 100 to 500 microgramsper meter squared body surface area (M2BSA), can enhance the immunereaction in aggressive tumors. Monocyte and lymphocyte activation isaugmented by INF-alpha, INF-beta and gamma. The IL-1 and IL-2 receptorantagonists are removed by ultrapheresis and thereby upregulate the invivo activity of these cytokines. An 80 kD glycoprotein, which isresponsible for inhibiting blastoid transformation in advancedmalignancy, chronic infectious disease and pregnancy, has recently beenfound, and appears to be responsible for the loss of delayedhypersensitivity reactions in these diseases, which is removed by thisprocess. This is significant because in removing this type ofsuppression, vaccines of all types will work better. Dosage regimes forIFN-alpha and beta are 3 M units subcutaneous three times a week up to20 M units/M2 BSA daily. Interferon-gamma is administered in a dosage ofbetween 100 to 1000 micgms per day.

D. Anti-Cytokine Receptor Molecules.

It is well established that TNF receptor 1 and TNF receptor 2 moleculesare shed by tumor cells, and that these molecules appear to inhibitimmune mediated attack by the host on the tumor cells. The ultrapheresisis believed to remove the majority of these soluble receptors.Additional, and/or selective, removal of these solublereceptor/inhibitors to soluble tissue necrosis factor receptor-1(“sTNFR-1”), soluble tissue necrosis factor receptor-2 (“sTNFR-2”),soluble interleukin-2 receptor (“sIL-2R”), soluble interleukin-1receptor (“sIL-1R”), soluble interleukin-6 receptor (“sIL-6R”), orsoluble interferon-gamma receptor (“sIFN-gammaR”) can be used tosupplement or instead of the ultrapheresis. The advantage of selectiveremoval is that the same beneficial effect is obtained in treatment ofthe disorder but the treatment is much less expensive and safer sinceexogenous plasma or albumin does not have to be administered to thepatient when there is selective removal. These can be removed by bindingto the cytokine, an epitope thereof, or an antibody to the receptor.These can be immobilized in the filter, in a column, or using otherstandard techniques for binding reactions to remove proteins from theblood or plasma of a patient. As used herein, antibody refers toantibody, or antibody fragments (single chain, recombinant, orhumanized), immunoreactive against the receptor molecules. In thepreferred embodiment, these antibodies are immobilized on theultrapheresis membrane filters, using standard antibody couplingtechniques. In the most preferred embodiment, the antibody is reactivewith the carboxy-terminus of the shed receptor molecules, thereby avoidconcerns with signal transduction by the receptor is still present onthe cell surface.

E. Chemotherapeutic Agents

Preferred chemotherapeutic agents are those which are synergistic withTNF, for example, alkylating agents, doxyrubicin, carboplatinum,cisplatinum, and tomoxifen. Tamoxifen plays a role not only in blockingof estrogen receptors but also certain growth factor receptors such asepidermal derived growth factor (“EDGF”), fibroblast derived growthfactor (“FDGF”), tumor derived growth factor (“TDGF”), TDGF-β andplatelet derived growth factor (“PDGF”) and therefore may becomplementary to inflammation against cancers provoked by ultrapheresis.

F. Radiation

Radiation therapy is destructive of normal tissue, causing tumors to diepartially by an inflammatory attack. Ultrapheresis allows the use oflower doses of radiation to kill residual tumor cells and spare normaltissue. In a preferred method, ultrapheresis is used as the initialtherapy, followed by radiation at approximately one-half of the normaldosages. It is well established that TNF kills tumor cells by generatingfree oxygen radicals, hydroxyl radicals and halide ions, and thatradiation therapy generates carbonium ions in tissue. Therefore thecombination of the two is more effective in killing cancer cells thaneither alone.

III. EXAMPLES Example 1 Treatment of a Patient with MetastaticLeiomyoscarcoma with Ultrapheresis

Mrs. J. K. is a 43 year old lady with metastatic leiomyoscarcoma withsix (6) lung metastases all of which developed within one month ofsurgery on both lungs to remove tumors. These tumors had also failedmethotrexate, adriamycin, ifosphomide, and dactinomycin.

She received 24 ultrapheresis procedures with no side effects. One monthlater, CAT scan revealed only four (4) tumors and these were reduced insize by 50%.

Example 2 Treatment of Patients with Breast Cancer by Ultrapheresis andThalidomide

Mrs. J. R. is a 44 year old lady who had metastatic breast cancer thathad failed radiation therapy and treatment with chemotherapeutic agents:cytoxan, adriamycin, 5-FU, taxol, cis-platin, navalbine, tamoxofin andarimedex. Tumor at the time of ultrapheresis was documented in lungs,bone and skin of the entire left anterior and lateral chest.

She was treated with 15 ultrapheresis procedures over a three weekperiod. She experienced marked inflammation in the tumors of his skin,increased pain from the tumors in her bones, and swelling of the tumorsin her lungs. She then received the drug thalidomide 200 mg at night.The redness and swelling in her skin improved within 2 days and herbreathing returned to normal within one week. Two weeks after completingtreatment, all tumor in her skin had resolved clinically, her bone painresolved and the tumors in her lungs resolved on repeat CAT scan. Oneweek later, she returned to work as a school counselor. She testeddisease free two months after treatment and was being maintained onthalidomide at the same dose.

Example 3 Treatment of Patient with Metastic Melanoma with Ultrapheresisand Thalidomide

Mr. P. G. is a 54 year old engineer with metastatic melanoma withmetastases to lung and to lymph nodes in the mediastinum.

He received 24 ultrapheresis procedures, resulting in a 25% reduction oftumors. He was subsequently treated with an additional 12 procedures,resulting in minor tumor reduction despite evidence of tumorinflammation. The tumors regrew within one month. He was again retreatedwith ultrapheresis, again resulting in inflammation and some minorregression, but was then treated with thalidomide at the time of tumorinflammation. Two months later, repeat CAT scan showed completedisappearance of tumors in the lung and mediastinum. He is beingfollowed closely and shows no evidence of disease and has no medicalcomplaints six months after completing treatment.

Example 4 Treatment of a Patient with Metastic Adenocarcinoma withUltrapheresis and Thalidomide

Dr. R. S. is a 59 year old gentlemen with metastatic adenocarcinoma ofthe left upper lung with metastases to liver, brain and bones. Histumors had failed to respond to taxol, cis-platin and etoposide. Hisbrain tumors had responded to radiation therapy.

He received 15 ultrapheresis procedures. Each procedure caused increasedpain in tumors of his spine, pelvis, right hip and left shoulder. Followup scans after ultrapheresis treatment revealed resolution of tumors inpelvis, spine, hip, and ribs. There was a 50% reduction in the primarytumor in the lung and liver. Thalidomide was then started at 200 mg eachnight. One month later, the scans revealed further reduction in thetumors in lung and liver. The patient's pains have all been resolved andhe is asymptomatic at this time.

Modifications and variations of the method and compositions describedherein will be obvious to those skilled in the art. Such modificationsand variations are intended to come within the scope of the appendedclaims.

1. A system for inducing an immune response against transformed,infected, or diseased tissue comprising a device for removing onlycomponents present in blood or plasma having a molecular weight of120,000 daltons or less, having an inlet and outlet for connection to apump and tubing to recirculate the blood or plasma of a patient throughthe device.
 2. A system according to claim 1, where said device hasimmobilized therein absorbents to remove specific cytokine or cellularinhibitors selectively from the blood, wherein said cytokine or cellularinhibitors are selected from the group consisting of soluble tumornecrosis factor receptor-1 (sTNFR-1), soluble tumor necrosis factorreceptor-2 (sTNFR-2), soluble interleukin-2 receptor (sIL-2R), solubleinterleukin-1 receptor (sIL-1R), soluble interleukin-6 receptor(sIL-6R), and soluble interferon-gamma receptor (sIFN-gammaR).
 3. Thesystem of claim 1 wherein the device is a capillary membrane filter witha pore size of between about 0.02 and 0.05 microns.
 4. The system ofclaim 1 wherein the device is a parallel plate filter with a pore sizeof between about 0.04 and 0.08 microns.
 5. The system of claim 1 whereinthe device comprises filters with different pore sizes or geometries toprovide for staggered removal of materials from the blood.
 6. The systemof claim 1 wherein the device is an absorbent column selectivelyremoving specific cytokine or cellular inhibitors from the blood orplasma.
 7. The system of claim 6 wherein the cytokine or cellularinhibitors are selected from the group consisting of sTNFR-1, sTNFR-2,sIL-2R, sIL-1R, sIL-6R, and sIFN-gammaR.
 8. The system of claim 7, wherethe absorbent column comprises cytokines or antibody or antibodyfragments immunoreactive with the cytokine or cellular inhibitors. 9.The system of claim 8 wherein the patient's blood or plasma iscirculated through the absorbent column prior to being returned to thepatient.
 10. The system of claim 1 further comprising a radiation sourceoperable to treat the patient.
 11. A method of inducing an immuneresponse against transformed, infected, or diseased tissue comprisingadministering to a patient blood, plasma, or a blood fraction from whichonly components having a molecular weight of 120,000 daltons or lesshave been removed, whereby administration of said blood, plasma, orblood fraction induces an immune response against transformed, infected,or diseased tissue in the patient until the transformed, infected, ordiseased tissue is reduced in amount.
 12. A method of inducing an immuneresponse against transformed, infected, or diseased tissue comprisingadministering to a patient blood, plasma, or a blood fraction from whichonly components having a molecular weight of 120,000 daltons or lesshave been removed by the system of claim 1, whereby administration ofsaid blood, plasma, or blood induces an immune response againsttransformed, infected, or diseased tissue in the patient until thetransformed, infected, or diseased tissue is reduced in amount.
 13. Themethod of claim 12 wherein the tissue is a solid tumor.
 14. The methodof claim 12 wherein the components are removed from one volume of bloodor plasma.
 15. The method of claim 12 wherein the components are removedin multiple treatments.
 16. The method of claim 12 wherein the tissue isto be treated further with an agent selected from the group consistingof anti-angiogenic compounds, procoagulant compounds, cytokines,chemotherapeutic agents, and radiation.
 17. The method of claim 16wherein the agent is a cytokine, and the cytokine is selected from thegroup consisting of GM-CSF, erythropoietin, thrombopoetin, G-CSF, M-CSF,and SCF.
 18. The method of claim 12 wherein the removal of thecomponents comprises selectively removing soluble cytokine receptormolecules.
 19. The method of claim 18 wherein the soluble cytokinereceptor molecules are removed by binding of the molecules to a filter.20. The method of claim 19 wherein the soluble cytokine receptormolecules are selected from the group consisting of sTNFR-1 and sTNFR-2.21. The method of claim 16 wherein the agent is thalidomide.
 22. Themethod of claim 12 wherein the patient is to be vaccinated with avaccine against the transformed, infected or diseased tissue.
 23. Amethod for preparing blood, plasma, or a blood fraction ex vivo forinducing an immune response against transformed, infected, or diseasedtissue until the transformed, infected, or diseased tissue is reduced inamount, the method comprising removing from provided blood or plasma exvivo only components having a molecular weight of 120,000 daltons orless.
 24. A method for inducing an immune response against transformed,infected, or diseased tissue in a patient by administering an agentselected from the group consisting of anti-angiogenic compounds,procoagulant compounds, cytokines, and chemotherapeutic agents to apatient treated with blood, plasma, or a blood fraction from which onlycomponents having a molecular weight of 120,000 or less have beenremoved.
 25. The method of claim 24 wherein the agent is a cytokine andthe cytokine is selected from the group consisting of GM-CSF,erythropoietin, thrombopoetin, G-CSF, M-CSF, and SCF.
 26. A kit fortreatment of a patient to induce an immune response against transformed,infected or diseased tissue comprising: a device for removing onlycomponents present in blood or plasma having a molecular weight of120,000 daltons or less and an agent selected from the group consistingof anti-angiogenic compounds, procoagulant compounds, cytokines,chemotherapeutic agents, and a radiation source, in a dosage formulationfor treatment of the patient.
 27. The kit of claim 26 wherein the agentis an anti-angiogenic compound.
 28. The kit of claim 26 wherein theagent is a procoagulant compound.
 29. The kit of claim 26 wherein theagent is a cytokine.
 30. The kit of claim 29 wherein the cytokine isselected from the group consisting of GM-CSF, erythropoietin,thrombopoetin, G-CSF, M-CSF, and SCF.
 31. The kit of claim 26 whereinthe agent is a chemo-therapeutic agent.
 32. The kit claim 31 wherein theagent is selected from the group consisting of alkylating agents,doxorubicin, carboplatin, cisplatin, and taxol.
 33. The kit of claim 26further comprising an anticoagulant to treat the device prior to use.34. The kit of claim 27 wherein the agent is thalidomide.